Feasibility analysis of inactivating influenza A(H1N1) virus using UVC robot in classroom environment.

Background: Starting from 2009, H1N1 has been one of the respiratory diseases that afflict the global population. Concurrently, due to the influence of COVID-19, it has become widely accepted that preventing the virus's spread necessitates personal protection measures and disinfection in public spaces. Experiments: This study conducted two experiments. In the classroom experiment, six UVC dose test points were calibrated to test whether the UVC dose at each testing point met the standards for inactivating IAVs and the time required to meet the standards. In the simulated classroom experiment, seven square slides made of IAVs were placed. After 10 min of robot movement, irradiated sterile square slides were made into suspension and injected into chicken embryos. Cultivate chicken embryos and conduct IAVs testing. Results: Classroom experiment has shown that 5 testing points can meet the standards for inactivating IAVs(3 mJ/cm2), with a required time of 80 min, 40 min, 15 min, 5 min and 10 min. The UVC dose for testing points that do not meet the standards in 80 min is only 0.5 mJ/cm2. The simulation classroom experiment outcomes revealed that 99.99 % of IAVs were deactivated. Furthermore, this study established both a desktop control group and a chair arm control group, both of which yielded identical results, indicating an inactivation logarithm of IAVs≥4log. Conclusion: The study presented that IAVs on the surface of an object can be effectively and rapidly deactivated at an irradiation density of 1.8 mW/cm2. Meanwhile, the study provides evidence of the feasibility of using the GXU robot to inactivate IAVs in a classroom environment.

Covalent Tethering of Cobalt Porphyrins on Phenolic Resins for Electrocatalytic Oxygen Reduction and Evolution Reactions.

The front cover artwork is provided by Rui Cao's group at Shaanxi Normal University. The image shows the design of Co-porphyrin-engineered phenolic resins with intramolecular phenolic hydroxyl groups to facilitate proton and electron transfers for efficient oxygen electrocatalysis, which is bioinspired by cytochrome c oxidases, and shows the excellent performance of Zn-air batteries assembled with the hybrid material. Read the full text of the Research Article at 10.1002/cphc.202400017.

Covalent Tethering of Cobalt Porphyrins on Phenolic Resins for Electrocatalytic Oxygen Reduction and Evolution Reactions.

Using functionalized supporting materials for the immobilization of molecular catalysts is an appealing strategy to improve the efficiency of molecular electrocatalysis. Herein, we report the covalent tethering of cobalt porphyrins on phenolic resins (PR) for improved electrocatalytic oxygen reduction reaction (ORR) and oxygen evolution reaction (OER). A cobalt porphyrin bearing an alkyl bromide substituent was covalently tethered on phenolic resins, through the substitution reaction of alkyl bromides with phenolic hydroxyl groups, to afford molecule-engineered phenolic resins (Co-PR). The resulted Co-PR was efficient for electrocatalytic ORR and OER by displaying an ORR half-wave potential of E1/2=0.78 V versus RHE and an OER overpotential of 420 mV to get 10 mA/cm2 current density. We propose that the many residual phenolic hydroxyl groups on PR will surround the tethered Co porphyrin and play critical roles in facilitating proton and electron transfers. Importantly, Co-PR outperformed unmodified PR and PR loaded with Co porphyrins through simple physical adsorption (termed Co@PR). The zinc-air battery assembled using Co-PR displayed a performance comparable to that using Pt/C+Ir/C. This work is significant to present phenolic resins as a functionalized material to support molecular electrocatalysts and demonstrate the strategy to improve molecular electrocatalysis with the use of phenolic resin residues.

Digital economy: an effective path for promoting residents' health in China.

The primary prerequisite for socioeconomic growth is good health, hence promoting residents' health is a vital objective of public policies. It is yet up for debate whether or not the digital economy (DE), which will be crucial to future economic growth, will eventually result in improvements in residents' health. Utilizing the China Family Panel Studies (CFPS) data in 2020, we explore how the DE affects residents' health. The findings reveal that residents' health is greatly enhanced by the DE. The eastern region sees a more dramatic improvement in residents' health as a result of the DE. Additionally, the DE can improve residents' health through the promotion of regional green development. The study's findings add to our knowledge of how the DE impacts residents' health while also offering recommendations for achieving universal health.

Spatial and temporal evolution and influencing factors of tourism eco-efficiency in Fujian province under the target of carbon peak.

Tourism eco-efficiency is an important indicator to measure the sustainable development of tourism with the core objective of "minimum resource input and environmental damage, maximum economic and social output". This paper explores the spatial and temporal evolution characteristics of tourism eco-efficiency and its influencing factors in Fujian Province from 2011 to 2020 by combining the unexpected output Super-SBM model, gravity center model, standard deviation ellipse, and optimal parameters-based geographical detector model. The results are as follows. Tourism eco-efficiency in Fujian Province showed an overall fluctuating growth trend with an increase of 160.065% during the study period. The coefficient of variation of tourism eco-efficiency tends to fluctuate and decrease, and regional differences narrow. The spatial differentiation is remarkable, and it presents the spatial distribution characteristics of "Xiamen > Nanping > Quanzhou > Fuzhou > Longyan > Ningde > Sanming > Zhangzhou > Putian". The spatial pattern of tourism eco-efficiency is obvious in the direction of "northeast-southwest", and the trajectory of the center of gravity is "southwest-northeast-northwest", with spatial characteristics tending towards agglomeration. Tourism industry structure has the greatest impact on tourism eco-efficiency, followed by education level, urbanization level, technology innovation level, infrastructure investment, and economic development level. This study provides reference suggestions for improving tourism eco-efficiency which helps to promote adequate, balanced, and sustainable development of the regional tourism industry.

Kinkéliba (Combretum micranthum) Leaf Extract Alleviates Skin Inflammation: In Vitro and In Vivo Study.

Kinkéliba (Combretum micranthum, Seh-Haw in Wolof) is a popular bush tea in West African countries. Although the kinkéliba plant's leaves have been widely consumed for its nutritional and medicinal properties, its benefits on skin health potential have been practically untouched. In human epidermal primary keratinocytes, vitexin and isovitexin-rich kinkéliba extract treatment significantly (p < 0.001) enhanced up to 39.6% of the cell survival rate decreased by UV radiation irritation. The treatment of kinkéliba leaf extracts also reduced the production of UV-induced pro-inflammatory cytokines IL-6 and IL-8 by 57.6% and 42.5%, respectively (p < 0.001), which cause skin redness and skin barrier dysfunction, as well as wrinkles and collagen degradation. The anti-inflammation efficacy of kinkéliba leaf extracts might involve significant inhibition on the levels of cellular reactive oxygen species (ROS) (-70.8%, p < 0.001) and nitrotyrosine (-56.9%, p < 0.05). Further topical applications of kinkéliba leaf extract gel were found to reduce sodium lauryl sulfate (SLS)-induced skin inflammation: at D7, the skin trans-epidermal water loss (TEWL) and skin redness (a* value) were both reduced by 59.81% (p < 0.001) and 22.4% (p < 0.001), compared with D0. In vitro and in vivo data support a new topical application of the kinkéliba leaf as an effective active ingredient for the treatment of skin inflammation, as well as subsequent barrier dysfunction and inflammaging.

Obesity promotes radioresistance through SERPINE1-mediated aggressiveness and DNA repair of triple-negative breast cancer.

Obesity is a risk factor in various types of cancer, including breast cancer. The disturbance of adipose tissue in obesity highly correlates with cancer progression and resistance to standard treatments such as chemo- and radio-therapies. In this study, in a syngeneic mouse model of triple-negative breast cancer (TNBC), diet-induced obesity (DIO) not only promoted tumor growth, but also reduced tumor response to radiotherapy. Serpine1 (Pai-1) was elevated in the circulation of obese mice and was enriched within tumor microenvironment. In vitro co-culture of human white adipocytes-conditioned medium (hAd-CM) with TNBC cells potentiated the aggressive phenotypes and radioresistance of TNBC cells. Moreover, inhibition of both cancer cell autonomous and non-autonomous SERPINE1 by either genetic or pharmacological strategy markedly dampened the aggressive phenotypes and radioresistance of TNBC cells. Mechanistically, we uncovered a previously unrecognized role of SERPINE1 in DNA damage response. Ionizing radiation-induced DNA double-strand breaks (DSBs) increased the expression of SERPINE1 in cancer cells in an ATM/ATR-dependent manner, and promoted nuclear localization of SERPINE1 to facilitate DSB repair. By analyzing public clinical datasets, higher SERPINE1 expression in TNBC correlated with patients' BMI as well as poor outcomes. Elevated SERPINE1 expression and nuclear localization were also observed in radioresistant breast cancer cells. Collectively, we reveal a link between obesity and radioresistance in TNBC and identify SERPINE1 to be a crucial factor mediating obesity-associated tumor radioresistance.

Identification of metabolites in different parts of Juandan Baihe (Lilium lancifolium) by UPLC-Q-TOF-MS and their hypoglycemic activities / 数字中医药（英文）

Objective @#To identify the main components in the extracts of different parts of Juandan Baihe (Lilium lancifolium) by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) technology and investigate their hypoglycemic activities.@*Methods@#The MS fragmentation pathways of the main types of compounds in Juandan Baihe (Lilium lancifolium) were studied, and the main components in the extracts were systematically identified using MS fragmentation pathways combined with MS mining technology. Based on the hyperglycemia male mouse model [specific pathogen free (SPF)-grade Kunming mice] induced by streptozotocin (intragastric administration of 80 mg/kg for 3 d), the hypoglycemic effects of extracts of Juandan Baihe (Lilium lancifolium) roots, stems, corms, leaves, and flowers were evaluated by measuring the changes of blood glucose, daily water consumption, daily food intake, and body weight.@*Result@#The MS fragmentation pathways of regalosides, dioscins, phenylpropanoids, flavonoids, and chlorogenic acids in Juandan Baihe (Lilium lancifolium) were clarified, and a mining method for compounds in this plant was constructed. A total of 58 compounds, including 6 chlorogenic acids, 14 regalosides, 13 phenylpropanoids, 5 flavonoids, and 20 dioscins, were identified from the roots, stems, corms, leaves, and flowers of Juandan Baihe (Lilium lancifolium). Among them, 30 compounds were reported for the first time from this plant. The root and corm extracts demonstrated significant hypoglycemic activities by reducing blood glucose levels from 23.76 ± 1.21 and 24.29 ± 1.35 mmol/L to 17.21 ± 1.23 and 18.78 ± 1.49 mmol/L, respectively (P < 0.05). The roots and corms extracts could also attenuate the symptoms of polydipsia (P < 0.01), polyphagia (P < 0.05), and weight loss caused by diabetes.@*Conclusion@#This study clarifies that the roots of Juandan Baihe (Lilium lancifolium) are rich in regalosides and dioscins for the first time, and have significant hypoglycemic activities, providing the foundation for the comprehensive utilization of this plant and the development of hypoglycemic drugs.

Slope climbing of urban expansion worldwide: Spatiotemporal characteristics, driving factors and implications for food security.

The tendency of global urban expansion to be slope climbing has partly become possible with scarce cropland resources in plains. However, the scientific understanding of the quantity, intensity, pattern, and effect of the slope climbing of urban expansion (SCE) is minimal globally. In this study, we have attempted to quantify and evaluate global SCE from Suomi National Polar-orbiting Partnership (SNPP)-Visible Infrared Imaging Radiometer Suite (VIIRS)-like data and other auxiliary data. Results revealed that global SCE areas unevenly increased from 22,760 km2 to 90,720 km2 from 2000 to 2020, with an annual growth rate of 21.72%, in which low-environment cost type areas increased from 21,550 km2 to 84,010 km2 while high-environment cost type (HEC) areas increased from 1210 km2 to 6710 km2. One remarkable phenomenon is that China's SCE areas in 2020 were more than 11 times those in 2000. In addition, global SCE intensity increased by about 3.4-fold from 2000 to 2020 and the rapid growth of HEC intensity is concentrated in Asia and North America. SCE is mostly affected by urban population growth and terrain. Economic development also promotes its development to a certain extent. We also noted that global SCE potentially made a considerable contribution to saved cropland, saving about 46,747 km2 with a theoretical increased grain yield of 25,020 × 103 t. Our study provides timely and transparent monitoring of global SCE and offers new insights into sustainable urban development.

Stressing the Regulatory Role of Long Non-Coding RNA in the Cellular Stress Response during Cancer Progression and Therapy.

Cellular stress response is an important adaptive mechanism for regulating cell fate decision when cells confront with stress. During tumorigenesis, tumor progression and the course of treatment, cellular stress signaling can activate subsequent response to deal with stress. Therefore, cellular stress response has impacts on the fate of tumor cells and tumor responsiveness relative to therapeutic agents. In recent years, attention has been drawn to long non-coding RNAs (lncRNAs), a novel class of RNA molecules with more than 200 nucleotides in length, which has little protein-coding potential and possesses various functions in multiple biological processes. Accumulating evidence has shown that lncRNAs are also engaged in the regulation of cellular stress response, particularly in cancers. Here, we summarize lncRNAs that have been reported in the adaptive response to major types of cellular stress including genotoxic, hypoxic, oxidative, metabolic and endoplasmic reticulum stress, all of which are often encountered by cancer cells. Specifically, the molecular mechanisms of how lncRNAs regulate cellular stress response during tumor progression or the development of therapy resistance are emphasized. The potential clinical applications of stress-responsive lncRNAs as biomarkers will also be discussed.

Amino acid restriction induces a long non-coding RNA UBA6-AS1 to regulate GCN2-mediated integrated stress response in breast cancer.

Oncogene activation, massive proliferation, and increased nutrient demands often result in nutrient and oxygen deprivation in solid tumors including breast cancer (BC), leading to the induction of oxidative stress and endoplasmic reticulum (ER) stress, and subsequently triggering integrated stress response (ISR). To elucidate the role of long non-coding RNAs (lncRNAs) in the ISR of BC, we performed transcriptome analyses and identified a lncRNA, UBA6-AS1, which was upregulated upon amino acid deprivation and ER stress. UBA6-AS1 was preferentially induced in triple-negative BC (TNBC) cells deprived of arginine or glutamine, two critical amino acids required for cancer cell growth, or treated with ER stress inducers. Mechanistically, UBA6-AS1 was regulated through the GCN2/eIF2α/ATF4 pathway, one of the major routes mediating ISR in amino acid sensing. In addition, both in vitro and in vivo assays indicated that UBA6-AS1 promoted TNBC cell survival when cells encountered metabolic stress, implicating a regulatory role of UBA6-AS1 in response to intratumoral metabolic stress during tumor progression. Moreover, PARP1 expression and activity were positively regulated by the GCN2/UBA6-AS1 axis upon amino acid deprivation. In conclusion, our data suggest that UBA6-AS1 is a novel lncRNA regulating ISR upon metabolic stress induction to promote TNBC cell survival. Furthermore, the GCN2-ATF4 axis is important for UBA6-AS1 induction to enhance PARP1 activity and could serve as a marker for the susceptibility of PARP inhibitors in TNBC.

Evaluation of clinical efficacy of suture-fixation mucopexy in the treatment of prolapsed hemorrhoid.

BACKGROUND: The aim of this study was to evaluate the clinical efficacy of suture-fixation mucopexy in the treatment of prolapsed hemorrhoids. METHODS: A total of 123 patients with grade II, III, and IV hemorrhoids were admitted to The TCM Hospital of Pu Dong New District between 2018 and 2019. They were randomly divided into the suture-fixation group (SF, N.=60) and the Milligan-Morgan hemorrhoidectomy group (MM, N.=63). Clinical efficacy, postoperative pain, average operation time, hospital stay, complications, and patient satisfaction were prospectively evaluated. RESULTS: No significant differences were identified in clinical efficacy; operation time and hospital stay between the two groups (P>0.05). However, VAS Score in the SF group was lower than that in the MM group. And the SF group was also more advantageous in anal function protection (P<0.05). In addition, the results of the follow-up survey revealed no significant difference in postoperative recurrence rate and patient satisfaction (P>0.05). CONCLUSIONS: Compared with Milligan-Morgan hemorrhoidectomy, suture-fixation mucopexy is as effective in the treatment of prolapsed hemorrhoid, but it has more advantages in reducing postoperative pain and protecting the anal function.

The Effects of Urban Forms on the PM2.5 Concentration in China: A Hierarchical Multiscale Analysis.

For a better environment and sustainable development of China, it is indispensable to unravel how urban forms (UF) affect the fine particulate matter (PM2.5) concentration. However, research in this area have not been updated consider multiscale and spatial heterogeneities, thus providing insufficient or incomplete results and analyses. In this study, UF at different scales were extracted and calculated from remote sensing land-use/cover data, and panel data models were then applied to analyze the connections between UF and PM2.5 concentration at the city and provincial scales. Our comparison and evaluation results showed that the PM2.5 concentration could be affected by the UF designations, with the largest patch index (LPI) and landscape shape index (LSI) the most influential at the provincial and city scales, respectively. The number of patches (NP) has a strong negative influence (-0.033) on the PM2.5 concentration at the provincial scale, but it was not statistically significant at the city scale. No significant impact of urban compactness on the PM2.5 concentration was found at the city scale. In terms of the eastern and central provinces, LPI imposed a weighty positive influence on PM2.5 concentration, but it did not exert a significant effect in the western provinces. In the western cities, if the urban layout were either irregular or scattered, exposure to high PM2.5 pollution levels would increase. This study reveals distinct ties of the different UF and PM2.5 concentration at the various scales and helps to determine the reasonable UF in different locations, aimed at reducing the PM2.5 concentration.

Covalent immobilization of the phytic acid-magnesium layer on titanium improves the osteogenic and antibacterial properties.

In order to improve early osseointegration and long-term survival rate of implants, a multifunctional titanium surface that promotes osteogenesis and antibacterial properties is expected. Incorporation of bioactive trace elements such as magnesium ions was proved a promising method to improve osseointegration of titanium. Phytic acid has strong chelating ability with multivalent cations, which has been used in surface modification. Moreover, phytic acid was proved antibacterial potential. Herein, to improve the osteogenic and antibacterial properties, a phytic acid-magnesium (PA-Mg) layer was introduced on titanium using phytic acid as a cross-linker molecule. No obvious changes of the surface characterization were observed by scanning electron microscopy and atomic force microscopy. X-ray photoelectron spectroscopy confirmed that the PA-Mg layer covalently bond to the Ti surface, and the thickness of the PA-Mg layer was about 150 nm. Besides, improved hydrophilic and more protein adsorption were observed on Ti-PA-Mg. Notably, a relatively controlled magnesium release was also observed on Ti-PA-Mg. Human bone mesenchymal stem cells showed better adhesion, proliferation, and osteogenic differentiation on Ti-PA-Mg samples, indicating improved biocompatibility and osteoinductivity. Moreover, Ti-PA-Mg had better antibacterial properties against porphyromonas gingivalis than Ti. Overall, the PA-Mg layer on Ti surface improved the osteogenic and antibacterial properties, which may have promise for use in dental implantation.

MMP-1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via the JNK and ERK pathway.

Enhancing the functions of mesenchymal stem cells (MSCs) is considered a potential approach for promoting tissue regeneration. In this study, we investigated the effects of Matrix Metalloproteinase-1 (MMP-1) on bone marrow mesenchymal stem cells (BMSCs) and its mechanism. Our results showed that knockdown of MMP-1 impeded scratch closure, attenuated proliferation, inhibited ALP activity, ALP denser staining and mineralization in vitro, and decreased expression of RUNX2, OSX, OPN and OCN in BMSCs, while 20 ng/mL recombinant human MMP-1 protein (rhMMP-1) significantly accelerated scratch closure, enhanced proliferation, ALP activity, ALP denser staining and mineralization in vitro, and increased expression of RUNX2, OSX, OPN and OCN. In addition, knockdown of MMP-1 inhibited the expression of phosphorylated c-Jun N-terminal kinase (p-JNK) and phosphorylated extracellular regulated protein kinases (p-ERK), while 20 ng/mL rhMMP-1 increased the expression of p-JNK and p-ERK in BMSCs. Furthermore, inhibition of c-Jun N-terminal kinase (JNK) and extracellular regulated protein kinases (ERK) by their inhibitor SP600125 and PD98059 dramatically blocked MMP-1-enhanced ALP activity and mineralization in BMSCs. Our results revealed that MMP-1 could accelerate the osteogenic differentiation potentials of BMSCs via the JNK and ERK pathway, providing the mechanism underlying MSC biology and identifying a potential target for improving bone tissue regeneration.

Hepatocellular carcinoma-derived high mobility group box 1 triggers M2 macrophage polarization via a TLR2/NOX2/autophagy axis.

In many human cancers, including hepatocellular carcinoma (HCC), high density of infiltrating tumor-associated macrophages (TAM) is associated with poor prognosis. Most TAMs express a M2 phenotype subsequently supporting tumor growth. How tumor cells polarize these TAMs to a pro-tumor M2 phenotype is still poorly understood. Our previous studies have revealed that a Toll-like receptor 2 (TLR2)-dependent autophagy triggered by hepatoma-derived factors down-regulates NF-κB p65 and drives M2 macrophage differentiation. However, the underlying mechanisms and potential hepatoma-derived TLR2 ligands are not clear. Here, we provide evidence to reveal that NADPH oxidase 2 (NOX2)-dependent reactive oxygen species (ROS) generation is crucial for HCC-induced autophagy, NF-κB p65 down-regulation and M2 phenotype polarization in primary macrophages. This NOX2-generated ROS production in abolished in TLR2-deficient macrophages. HCC-derived or recombinant high-mobility group box 1 (HMGB1) is able to trigger this TLR2-mediated M2 macrophage polarization. Blockage of HMGB1 and ROS by inhibitors, ethyl pyruvate and N-acetylcysteine amide, respectively, significantly reduces both M2 macrophage accumulation and liver nodule formation in HCC-bearing mice. Our findings uncover a HMGB1/TLR2/NOX2/autophagy axis to trigger M2 macrophage polarization in HCC that can be considered as a novel therapeutic target for treating HCC.

Does China's Urban Development Satisfy Zipf's Law? A Multiscale Perspective from the NPP-VIIRS Nighttime Light Data.

Currently, whether the urban development in China satisfies Zipf's law across different scales is still unclear. Thus, this study attempted to explore whether China's urban development satisfies Zipf's law across different scales from the National Polar-Orbiting Partnership's Visible Infrared Imaging Radiometer Suite (NPP-VIIRS) nighttime light data. First, the NPP-VIIRS data were corrected. Then, based on the Zipf law model, the corrected NPP-VIIRS data were used to evaluate China's urban development at multiple scales. The results showed that the corrected NPP-VIIRS data could effectively reflect the state of urban development in China. Additionally, the Zipf index (q) values, which could express the degree of urban development, decreased from 2012 to 2018 overall in all provinces, prefectures, and counties. Since the value of q was relatively close to 1 with an R2 value > 0.70, the development of the provinces and prefectures was close to the ideal Zipf's law state. In all counties, q > 1 with an R2 value > 0.70, which showed that the primate county had a relatively stronger monopoly capacity. When the value of q < 1 with a continuous declination in the top 2000 counties, the top 250 prefectures, and the top 20 provinces in equilibrium, there was little difference in the scale of development at the multiscale level with an R2 > 0.90. The results enriched our understanding of urban development in terms of Zipf's law and had valuable implications for relevant decision-makers and stakeholders.

A low-cost, automated parasite diagnostic system via a portable, robotic microscope and deep learning.

Manual hand counting of parasites in fecal samples requires costly components and substantial expertise, limiting its use in resource-constrained settings and encouraging overuse of prophylactic medication. To address this issue, a cost-effective, automated parasite diagnostic system that does not require special sample preparation or a trained user was developed. It is composed of an inexpensive (~US$350), portable, robotic microscope that can scan over the size of an entire McMaster chamber (100 mm2 ) and capture high-resolution (~1 µm lateral resolution) bright field images without need for user intervention. Fecal samples prepared using the McMaster flotation method were imaged, with the imaging region comprising the entire McMaster chamber. These images are then automatically segmented and analyzed using a trained convolution neural network (CNN) to robustly separate eggs from background debris. Simple postprocessing of the CNN output yields both egg species and egg counts. The system was validated by comparing accuracy with hand-counts by a trained operator, with excellent performance. As a further demonstration of utility, the system was used to conveniently quantify drug response over time in a single animal, showing residual disease due to Anthelmintic resistance after 2 weeks.

[Toxoplasma gondii ROP38 promotes the maturation of dendritic cells mediated by TLR4].

Objective To investigate the effect of rhoptry protein 38 (ROP38) from Toxoplasma gondii (T. gondii) on the maturation of dendritic cells (DCs) by Toll-like receptor 4 (TLR4) induction in vitro. Methods The total RNA from T. gondii RH strain was extracted by guanidine thiocyanate method, and then cDNA was synthesized with reverse transcription reaction. After ROP38 gene was amplified by PCR, the recombinant pGEX-4T-ROP38 was constructed and expressed under IPTG induction. The recombinant ROP38 protein was detected by SDS-PAGE and Western blot analysis. The secondary structure and antigenicity of the ROP38 were predicted through DNAStar8.0 and ProtScale. In vitro, DCs were isolated and cultured for 6 days, then reacted with ROP38 antigen for 2 hours. The CD11c was detected by flow cytometry, and the data were analyzed with ANOVA by SPSS 23.0 software. Results The amplified gene was about 516 bp as expected. The sequence analysis showed that its homology was 99% compared with the reported sequence (XM_002366710.2) from GenBank. It was found that the relative molecular mass (Mr) of recombinant ROP38 was 45 kD. The prediction of structure indicated that there were 5 α-helices, 5 ß-sheets, 5 hydrophilic regions and 8 potential epitopes in ROP38 protein. In vitro, the expression of CD11c on DCs was significantly up-regulated after stimulated with ROP38, and the expression level of CD11c in ROP38 infection group was significantly higher than that of the other groups. Conclusion ROP38 promotes the maturation of DCs mediated by TLR4.

Natural variation in the promoter of rice calcineurin B-like protein10 (OsCBL10) affects flooding tolerance during seed germination among rice subspecies.

Rice (Oryza sativa L.) has two ecotypes, upland and lowland rice, that have been observed to show different tolerance levels under flooding stress. In this study, two rice cultivars, upland (Up221, flooding-intolerant) and lowland (Low88, flooding-tolerant), were initially used to study their molecular mechanisms in response to flooding germination. We observed that variations in the OsCBL10 promoter sequences in these two cultivars might contribute to this divergence in flooding tolerance. Further analysis using another eight rice cultivars revealed that the OsCBL10 promoter could be classified as either a flooding-tolerant type (T-type) or a flooding-intolerant type (I-type). The OsCBL10 T-type promoter only existed in japonica lowland cultivars, whereas the OsCBL10 I-type promoter existed in japonica upland, indica upland and indica lowland cultivars. Flooding-tolerant rice cultivars containing the OsCBL10 T-type promoter have shown lower Ca2+ flow and higher α-amylase activities in comparison to those in flooding-intolerant cultivars. Furthermore, the OsCBL10 overexpression lines were sensitive to both flooding and hypoxic treatments during rice germination with enhanced Ca2+ flow in comparison to wild-type. Subsequent findings also indicate that OsCBL10 may affect OsCIPK15 protein abundance and its downstream pathways. In summary, our results suggest that the adaptation to flooding stress during rice germination is associated with two different OsCBL10 promoters, which in turn affect OsCBL10 expression in different cultivars and negatively affect OsCIPK15 protein accumulation and its downstream cascade.